3.Establishment of model of diabetes and lens posterior capsule opacification induced by alloxan in rabbit
Qi, WEI ; Jin-mao, CHEN ; Min-li, HUANG ; Xia, LI ; Jian-feng, HE ; Shao-jian, TAN
Chinese Journal of Experimental Ophthalmology 2011;29(2):130-134
Background The establishment of diabetic animal model is a crucial step for the study about diabetic eye diseases. At present,the main modeling method include the injection of streptozotocin and alloxan. But the shortcoming of the former is an expensive price, and that of the later is high death rate of animals. Objective This experiment was to discuss the way which decrease the death of alloxan-injected animal and explore the effects of high blood glucose on the posterior capsular opacification (PCO). Methods Forty clean healthy male New Zealand white rabbits were randomly divided into 2 groups. 90mg/kg of alloxan were injected via ear vein once in 20 rabbits to create the diabetic animal models,and the equivalent amount of normal saline solution was injected at the same way as normal blood glucose group. The successful models were selected in the animals with the blood glucose level over 12. 0 mmol/L two weeks later, and PCO of lens were graded based on the method of Odrieh under the slit lamp. Extracapsular lens extraction was then performed on the right eye of rabbits in both groups, and the posterior capsules were obtained from these eyes at the 6th, 10th and 14th days after operation. The expression of proliferating cell nuclear antigen ( PCNA ) in posterior capsular lens epithelial cell was detected by immunohistochemistry. Results The modeling successful rate was 70% after injection of alloxan. The body weight of rabbits in high blood glucose group was significantly lowed and the blood glucose was significantly elevated in comparison with normal blood glucose group ( all P<0. 05). Two weeks after surgery ,2 eyes occurred 2 grade of PCO and only one eye showed the 1 grade of PCO in the high blood glucose group. However, 1 grade of PCO was found in 3 eyes in the normal blood glucose group. Biopsy revealed that PCNA was positively expressed in the cell nuclei of LECs in high blood glucose group rather than the normal blood glucose group from the 10th day after surgery. The proliferation index of PCNA was 0. 86±0. 04 and 0. 25±0. 03 respectively in high blood glucose group and normal blood glucose group, showing a significant difference between them (t = -16. 171 ,P = 0. 000). Conclusion Stable diabetic models of rabbits can be created by intravenous injection of 90 mg/kg alloxan. High blood glucose level is one of the important factors for the development of PCO.
4.ABO-incompatible allogeneic hematopoietic stem cell transplantation for treating malignant hematologic diseases
Xiaoyan SHAO ; Jian OUYANG ; Bing CHEN ; Dingyan XU ; Yonggong YANG ; Min ZHOU
Chinese Journal of Tissue Engineering Research 2008;12(12):2383-2386
BACKGROUND:ABO-incompatibility between donor and recipient is not a barrier for Successful allogeneic hematopoietic stem cell transplantation even though it is well established that major ABO incompatibility may lead to prolonged destruction of donor-derived erythrocytos and prolonged transfusioil requirements.OBJECTIVE:To explore the effect of ABO.incompatible on clinical characteristics in allogeneic-hematopoietic stem cell transplantation.DESIGN:A retrospective observation.SETTING:Department of Hematology.the Affiliated Drum Tower Hospital of Nanjing University Medical School.PARTICIPANTS:Fourteen patients(11 males and 3 feiliales,aged 15-60 years old)with malignant hematologic diseases who received ABO-incompatible allogeneic hematopoietic stem cell transplantation in the Affiliated Drum Tower Hospital of Nanjing University Medical School from May 2002 to September 2007 Were recruited for this study.Of the 14 patients,7 were human leukocyte antigen(HLA).matched,and the other 7 were HLA-half-matched.Controls were 11 patients who received ABO-compatibility bone marrow transplantation during the same period.Written informed consents for receiving allogeneic hematopoietic stem cell transplantation were obtained from each reciplent.The donors were sibling sister,sibling brother.son and mother,and they all agreed to provide marrow for transplantation.T1lis experiment was given an approval by the Ethics Committees of the hospital.METHODS:Regimen conditioning:HLA-matched transplantation regimen conditioning consisted of busulfan(Bu)and cyclophosphamide(Cy).HLA-half-matched transplantation regimen conditioning adopted GIAC program from Beijing People's Hospital.The GIAC program consisted of 4 parts:G:granulocyte colony-stimulating factors used for donors;I:stronger immunosuppressive regimen conditioning used for recipients;A: antihuman thymocyte globulin added:C: combined transplantation of bone marrow and peripheral blood;Perfusion of hematopoietic stem cells:The marrow from ABO-incompatible donor depleted erythrocytes by hydroxyethyl starch sedimentation.MAIN OUTCOME MEASURES:Adverse reaction.complication and hematologic recovery after ABO-incompatibility stem cell transplantation.RESULTS:One out of fourteen recipients developed pure red cell aplasia(PRCA)and dropped out of final analysis.Hematologic recovery:The median time of erythrocyte recovery after ABO-incompatible stem cell transplantation was delayed compared with ABO compatible stem cell transplantation (t=2.352.P<0.05).There were no significant difieFences in the recovery of neutrophils and platelets between ABO-incompatible group and ABO-compatible group(P>0.05).The median time of recovery of the erythrocyte and the blood type switching was delayed in HLA-mis-matched allogeneic hematopoietic stem cell transplantation compared with HLA-matched allogeneic hematopoietic stem cell transplantation,but without significant difference(P>0.05).Complications:During the stem cell transfusion following transplantation.none of 14 Patieats had hemolytic complications or delayed haemolysis.CONCLUSION:There was no evidence of ABO-incompatibility between donor and recipient is a barrier for successful allogeneic hematopoietic stem cell transplantation.
6.Bioinformatic analysis of small RNA expression during chondrogenesis in rats
Shibin SHAO ; Zixin MIN ; Yuanxu GUO ; Quancheng WANG ; Mengyao SUN ; Yan HAN ; Jian SUN
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(4):462-466
Objective To study the profiles and function of small RNA (sRNA)gene during chondrogenesis in rats so as to clarify the mechanisms of chondrocytes proliferation and differentiation.Methods All the sRNAs were identified from the female SD rats femoral head cartilages at three time points:at birth,ablactation and maturation,and three sRNA libraries were constructed.The Solexa sequencing and the bioinformatics analysis were employed to be blasted with the genomes of SD rats.Results The perfect match reads in the three libraries were screened out,which were correspondent to the 21 7 921 (41.23%),1 96 650 (38.74%)and 245 436 (41.54%)unique sRNA sequence,respectively.The percentages of 20-24 nt sRNA were 71.94% (d0),72.85% (d21),and 86.39%(d42).Half of clean sequences were 22 nt sRNA.The distribution characteristics of the reads were in line with the high-quality sRNA.More than 62% clean reads were from mature miRNA while the ratios in the three libraries were only 0.69%,0.78% and 0.63%.About 60% of the unique sRNA could not be matched with miRBase20.0 or Rfam9.1.Conclusion The distribution model of miRNA in the three libraries indicates that the miRNAs with different functions or from different sources are involved in the regulation of chondrocytes proliferation and differentiation in bone development and formation.
7.Cloning, over-expression and in vitro activity of aminoglycoside phosphotransferase from clinical MRSA isolates
Junsheng CHEN ; Min SU ; Daijie CHEN ; Jian LI ; Shidong KAN ; Lei SHAO
Journal of China Pharmaceutical University 2010;41(1):81-85
Aim: To clone and over-express the gene encoding aminoglycoside(AG)phosphotransferase(APH)from clinical MRSA isolates in E.coli and to develop an assay method for the recombinant APH.Methods: The susceptibility of clinical MRSA isolates to AGs was tested by disk diffusion.A nucleic acid sequence encoding APH was amplified from the genomic DNA of an isolate and ligated to expression vector pET-28a,and then trans-formed into E.coli BL21(DE3).After purification of the recombinant protein by affinity chromatography,the phosphorylation activity of the enzyme was determined by ESI-MS and disk diffusion.Flesults: All 6 clinical MRSA isolates were unsusceptible to AGs.After cloning and expression,the recombinant APH was purified to90%.The in vitro activity assay indicated that the recombinant protein could inactivate kanamycin B in the assay mixture within 2 h.Conclusion: The recombinant APH showed excellent enzymatic activity.The assay method was simple and convenient,which may provide the basis of developing a screening model for APH inhibitors.
8.Influence of ischemia/reperfusion on function of vascular endothelial cells and effect of intervention with drug
Chang-Lai ZHANG ; Jian-Li WANG ; Hui-Min SONG ; Jun SHAO ; Li-Hong LV ;
Chinese Journal of Current Advances in General Surgery 2004;0(05):-
Objective:To explore the influence of ischemia/reperfusion (anoxia/reoxygenation)[FK(W16*2。142mmZQ1mm]on immunofunction of endothelial cells(ECs) and effect of intervention with 2-hydroxymethyl-3,5,6-trimethylpyrazine on it.Methods:Model of ECs induced by anoxia/reoxygenation was established to mimic ECs ischemia/reperfusion injury in vivo with human umbilical vein endothelial cell line ECV304.Then 2-hydroxymethyl-3,5,6-trimethylpyraxine was used to intervene the anoxia/reoxygenation process.Nuclear transcriptional factor-?B(NF-?B) was exhibited by fluorescent staining,HLA-ABC,HLA-DR,CD86 and CD54 were detected by flow cytometry.Mixed endothelial cell-lymphocyte reaction(MELR) was conducted to examine the proliferation of lymphocyte,production of IL-2 and percentage of apoptotic lymphocyte.Results:Anoxia/reoxygenation made the ECV304 cell became round,shrunk and abscised,with increased plasma NF-?B,and changed from positive cytoplasm to positive nucleus.HLA-ABC,HLA-DR and CD86 on surface of cells increased but CD54 showed unchanged.MELR showed the incorporation of ~3H-TdR and production of IL-2 increased significantly and the percentage of apoptotic lymphocyte decreased.After 2-hydroxymethyl-3,5,6-trimethylpyrazine intervention,the ECV304 cell shapes recovered,NF-?B expression didn’t down-regulated,but the percentage of positive cells decreased,changes to positve dominant.Besides,reversal changes were shown in other parameters.Conclusion:Anoxia/reoxygenation influences some important immune related molecules in ECV304 cells.2-hydroxymethyl-3,5,6-trimethylpyrazine could antagonize these influences to maintain the immune function of endothelial cells in a relative normal manner.
9.Isolation and Identification on a Thermoacidophilic Fungus of High-producing Cellulase and the Characteristics of its Enzyme
Jian-Min GAO ; Hai-Bo WENG ; Yu XI ; Ming-Xue YUAN ; Shao-Yin HAN ;
Microbiology 1992;0(04):-
A novel Aspergillus terreus strain M11 was isolated from the compost containing cellulose and identified. The isolate grow best at 45℃ and pH2.0. It was found that the activity of the CMCase was up to 3.680IU/mL with high heat stability and the optimal reaction conditions of the CMCase were at 60℃ and pH2.0.
10.Correlation study of cerebral microbleeds with chronic renal failure in patients with acute lacunar stroke
Houqin CHEN ; Min HE ; Wenbing WANG ; Lingyun SHAO ; Jian ZUO ; Fei SONG
Journal of Medical Postgraduates 2017;30(3):294-297
Objective Cerebral small vessel disease is closely related to kidney disease .Chronic kidney disease ( CKD) may increase the risk of hemorrhage stroke .However, its impact on hemorrhage-prone small vessel disease represented by cerebral microb-leeds( CMBs) remains unclear .The purpose of this study was to investigate the relationship of CKD with the presence and location of CMBs in patients with acute lacunar stroke . Method Consecutive patients with acute lacunar stroke within 7 days from onset were enrolled retrospectively from January 2014 to July 2016 and scanned by gradient-echo T2*-weighted imaging (GRE-T2*WI).Their demographic, clinical, laboratory and imaging data were collected .Estimate glomerular filtration rate (eGFR) was calculated individu-ally by the following chronic kidney disease epidemiology collaboration (CKD-EPI) equation for the Asian population .CKD was defined as the level of eGFR<60 mL/min/1.73 m2. Results Finally, 308 patients (mean age:65.79±8.67 years; median NHISS:3(2-5);42.2%Female) with lacunar ischemic stroke were enrolled in the final analysis .Among these patients, CMBs were present in 116 patients ( 37.7%) and CKD in 62 patients ( 20.1%) .Patients were divided into CKD group and normal group according to GFR level . The result of univariate analysis showed that patients with CKD had higher prevalence of diabetes ( P=0.014) and higher degrees of CMBs (P=0.001) compared with normal group.CMBs were refined by its location .The result of multivariable analysis showed that CMBs in deep brain [ OR=7.61, 95%CI 4.18-16.55, P=0.001] were sig-nificantly associated with CKD incidence , while no significant rela-tionship was found in CKD incidence and CMBs in the lobe and mixed location of brain . Conclusion The CKD incidence in patients with acute lacunar stroke is in dependent relationship with CMBs in deep brain and without significant correlation with CMBs in the lobe and mixed location of brain .